CN108872217A - The synthesis and application of iridium dioxide nano enzyme - Google Patents
The synthesis and application of iridium dioxide nano enzyme Download PDFInfo
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- CN108872217A CN108872217A CN201810583228.6A CN201810583228A CN108872217A CN 108872217 A CN108872217 A CN 108872217A CN 201810583228 A CN201810583228 A CN 201810583228A CN 108872217 A CN108872217 A CN 108872217A
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- nano enzyme
- sarcosine
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- 102000004190 Enzymes Human genes 0.000 title claims abstract description 66
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 66
- HTXDPTMKBJXEOW-UHFFFAOYSA-N dioxoiridium Chemical compound O=[Ir]=O HTXDPTMKBJXEOW-UHFFFAOYSA-N 0.000 title claims abstract description 43
- 230000015572 biosynthetic process Effects 0.000 title claims abstract description 10
- 238000003786 synthesis reaction Methods 0.000 title claims abstract description 10
- FSYKKLYZXJSNPZ-UHFFFAOYSA-N sarcosine Chemical compound C[NH2+]CC([O-])=O FSYKKLYZXJSNPZ-UHFFFAOYSA-N 0.000 claims abstract description 66
- 108010077895 Sarcosine Proteins 0.000 claims abstract description 33
- 229940043230 sarcosine Drugs 0.000 claims abstract description 32
- 102000003992 Peroxidases Human genes 0.000 claims abstract description 22
- 108040007629 peroxidase activity proteins Proteins 0.000 claims abstract description 22
- 230000000694 effects Effects 0.000 claims abstract description 17
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 10
- 108010060059 Sarcosine Oxidase Proteins 0.000 claims abstract description 8
- 102000008118 Sarcosine oxidase Human genes 0.000 claims abstract description 8
- 238000004445 quantitative analysis Methods 0.000 claims abstract description 5
- 238000001514 detection method Methods 0.000 claims description 22
- 239000000243 solution Substances 0.000 claims description 15
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- 230000031700 light absorption Effects 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
- 239000008351 acetate buffer Substances 0.000 claims description 7
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 5
- 239000001301 oxygen Substances 0.000 claims description 5
- 229910052760 oxygen Inorganic materials 0.000 claims description 5
- 239000008363 phosphate buffer Substances 0.000 claims description 5
- 230000008569 process Effects 0.000 claims description 5
- 239000006228 supernatant Substances 0.000 claims description 4
- -1 Diamine salts Chemical class 0.000 claims description 2
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 claims description 2
- 238000001816 cooling Methods 0.000 claims description 2
- CEYULKASIQJZGP-UHFFFAOYSA-L disodium;2-(carboxymethyl)-2-hydroxybutanedioate Chemical compound [Na+].[Na+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O CEYULKASIQJZGP-UHFFFAOYSA-L 0.000 claims description 2
- 238000006243 chemical reaction Methods 0.000 abstract description 8
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 abstract description 6
- 239000003054 catalyst Substances 0.000 abstract description 4
- 230000002378 acidificating effect Effects 0.000 abstract description 2
- 239000003814 drug Substances 0.000 abstract 1
- 238000000855 fermentation Methods 0.000 abstract 1
- 230000004151 fermentation Effects 0.000 abstract 1
- 239000000126 substance Substances 0.000 abstract 1
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 12
- 239000000463 material Substances 0.000 description 10
- 239000004471 Glycine Substances 0.000 description 6
- DDRJAANPRJIHGJ-UHFFFAOYSA-N creatinine Chemical compound CN1CC(=O)NC1=N DDRJAANPRJIHGJ-UHFFFAOYSA-N 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 6
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 4
- 206010028980 Neoplasm Diseases 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- 239000002086 nanomaterial Substances 0.000 description 4
- 239000002105 nanoparticle Substances 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 229910052709 silver Inorganic materials 0.000 description 4
- 238000006555 catalytic reaction Methods 0.000 description 3
- 229940109239 creatinine Drugs 0.000 description 3
- 210000004907 gland Anatomy 0.000 description 3
- 229910052737 gold Inorganic materials 0.000 description 3
- 210000002700 urine Anatomy 0.000 description 3
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical compound N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- BAVYZALUXZFZLV-UHFFFAOYSA-N Methylamine Chemical compound NC BAVYZALUXZFZLV-UHFFFAOYSA-N 0.000 description 2
- 206010060862 Prostate cancer Diseases 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 229960001231 choline Drugs 0.000 description 2
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 2
- CVSVTCORWBXHQV-UHFFFAOYSA-N creatine Chemical compound NC(=[NH2+])N(C)CC([O-])=O CVSVTCORWBXHQV-UHFFFAOYSA-N 0.000 description 2
- SZVJSHCCFOBDDC-UHFFFAOYSA-N ferrosoferric oxide Chemical compound O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 description 2
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000003907 kidney function Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- NUJOXMJBOLGQSY-UHFFFAOYSA-N manganese dioxide Chemical compound O=[Mn]=O NUJOXMJBOLGQSY-UHFFFAOYSA-N 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 1
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 229910002902 BiFeO3 Inorganic materials 0.000 description 1
- 229910020197 CePO4 Inorganic materials 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 229910003321 CoFe Inorganic materials 0.000 description 1
- 229910002518 CoFe2O4 Inorganic materials 0.000 description 1
- 229910005335 FePt Inorganic materials 0.000 description 1
- 102000003983 Flavoproteins Human genes 0.000 description 1
- 108010057573 Flavoproteins Proteins 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- KEAYESYHFKHZAL-UHFFFAOYSA-N Sodium Chemical compound [Na] KEAYESYHFKHZAL-UHFFFAOYSA-N 0.000 description 1
- 102000019197 Superoxide Dismutase Human genes 0.000 description 1
- 108010012715 Superoxide dismutase Proteins 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000003990 capacitor Substances 0.000 description 1
- CETPSERCERDGAM-UHFFFAOYSA-N ceric oxide Chemical compound O=[Ce]=O CETPSERCERDGAM-UHFFFAOYSA-N 0.000 description 1
- 229910000422 cerium(IV) oxide Inorganic materials 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- UBEWDCMIDFGDOO-UHFFFAOYSA-N cobalt(II,III) oxide Inorganic materials [O-2].[O-2].[O-2].[O-2].[Co+2].[Co+3].[Co+3] UBEWDCMIDFGDOO-UHFFFAOYSA-N 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 229910000153 copper(II) phosphate Inorganic materials 0.000 description 1
- 229960003624 creatine Drugs 0.000 description 1
- 239000006046 creatine Substances 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- OCLWHYOULMTOJB-UHFFFAOYSA-L disodium hydrogen carbonate hydrate Chemical compound O.[Na+].[Na+].OC(O)=O.[O-]C([O-])=O OCLWHYOULMTOJB-UHFFFAOYSA-L 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- WBJZTOZJJYAKHQ-UHFFFAOYSA-K iron(3+) phosphate Chemical compound [Fe+3].[O-]P([O-])([O-])=O WBJZTOZJJYAKHQ-UHFFFAOYSA-K 0.000 description 1
- 229910000399 iron(III) phosphate Inorganic materials 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 229910044991 metal oxide Inorganic materials 0.000 description 1
- 150000004706 metal oxides Chemical class 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 229910052961 molybdenite Inorganic materials 0.000 description 1
- CWQXQMHSOZUFJS-UHFFFAOYSA-N molybdenum disulfide Chemical compound S=[Mo]=S CWQXQMHSOZUFJS-UHFFFAOYSA-N 0.000 description 1
- 229910052982 molybdenum disulfide Inorganic materials 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 229910000510 noble metal Inorganic materials 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- SIWVEOZUMHYXCS-UHFFFAOYSA-N oxo(oxoyttriooxy)yttrium Chemical compound O=[Y]O[Y]=O SIWVEOZUMHYXCS-UHFFFAOYSA-N 0.000 description 1
- 239000006174 pH buffer Substances 0.000 description 1
- 150000002978 peroxides Chemical class 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- BHZOKUMUHVTPBX-UHFFFAOYSA-M sodium acetic acid acetate Chemical compound [Na+].CC(O)=O.CC([O-])=O BHZOKUMUHVTPBX-UHFFFAOYSA-M 0.000 description 1
- FDRCDNZGSXJAFP-UHFFFAOYSA-M sodium chloroacetate Chemical compound [Na+].[O-]C(=O)CCl FDRCDNZGSXJAFP-UHFFFAOYSA-M 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 229910006297 γ-Fe2O3 Inorganic materials 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J23/00—Catalysts comprising metals or metal oxides or hydroxides, not provided for in group B01J21/00
- B01J23/38—Catalysts comprising metals or metal oxides or hydroxides, not provided for in group B01J21/00 of noble metals
- B01J23/40—Catalysts comprising metals or metal oxides or hydroxides, not provided for in group B01J21/00 of noble metals of the platinum group metals
- B01J23/46—Ruthenium, rhodium, osmium or iridium
- B01J23/468—Iridium
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- B01J35/00—Catalysts, in general, characterised by their form or physical properties
- B01J35/20—Catalysts, in general, characterised by their form or physical properties characterised by their non-solid state
- B01J35/23—Catalysts, in general, characterised by their form or physical properties characterised by their non-solid state in a colloidal state
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- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01G—COMPOUNDS CONTAINING METALS NOT COVERED BY SUBCLASSES C01D OR C01F
- C01G55/00—Compounds of ruthenium, rhodium, palladium, osmium, iridium, or platinum
- C01G55/004—Oxides; Hydroxides
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N31/00—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/535—Production of labelled immunochemicals with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/70—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving creatine or creatinine
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/34—Genitourinary disorders
- G01N2800/347—Renal failures; Glomerular diseases; Tubulointerstitial diseases, e.g. nephritic syndrome, glomerulonephritis; Renovascular diseases, e.g. renal artery occlusion, nephropathy
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Abstract
The present invention relates to nano-catalytics, analytical chemistry field, specifically include iridium dioxide（IrO2）The synthesis and application of nano enzyme.IrO2Nano enzyme has the activity of peroxidase at acidic, can catalyzing hydrogen peroxide and organicvisualization reagent generation chromogenic reaction.The present invention utilizes the IrO being prepared2Nano enzyme and sarcosine oxidase are coupled the quantitative analysis to realize sarcosine.The invention can be used for food fermentation, biological medicine, chemical industry, environment, the quantitative analysis of biotechnology field and catalyst etc..
Description
Technical field
The present invention relates to analytical chemistry fields, specifically include iridium dioxide（IrO2）The imitative enzyme material of its conduct of the synthesis of nano enzyme
The application of material.
Background technique
Nano enzyme is the special performance of a kind of existing nano material, and has the analogue enztme of catalysis.Nano enzyme overcomes
Many disadvantages of native enzyme, such as expensive, easy in inactivation and condition of storage require harshness, to bio-sensing, immunoassay, cancer
The fields such as disease diagnosing and treating produce tremendous influence.
Up to the present, a variety of inorganic nano materials all show imitative peroxidase, imitative oxidizing ferment, imitative hydrogen peroxide
The performances such as enzyme or imitative superoxide dismutase.Wherein, imitative peroxidase activity refers to that nano material has with peroxidase one
Sample effect with H2O2For the enzyme of electron acceptor catalysis substrate oxidation.
The reported inorganic nano material with imitative peroxidase activity includes Fe3O4、γ-Fe2O3、FePO4、
FePt、CuO、Cu3(PO4)2、CuS、Cu、CuInS2、Co3O4、CoFe、CoFe2O4、CeO2、CePO4、Gd、MnO2、MnSe、ZnO、
BiFeO3、MoS2 、WC、VO2、V2O3、Ir、Pd-Ir、Au、Pt、Au/CuS、Bi/Au、Ag/Pt、Ag/Pd、Ag/Au、Ag/Pt、
Au/Pd、Si-dots、V2O5.It is worth noting that, not all metal oxide all has imitative peroxidase activity.Two
Yttrium oxide has not been reported with Mimicry enzyme.
Noble metal and its oxide nano particles have high catalytic activity to some chemical reactions.IrO2With many
Important application, including as super capacitor and electrochromic material, stimulating electrode between neuron, what is sensed and be imaged for pH is super
Microelectrode and the catalyst reacted for oxygen and chlorine.IrO2Nano particle is interior to water decomposition oxygen evolution reaction at a wider pH range
Show higher electro catalytic activity.IrO2Nano particle used also as the electronics conductive matrix of enzyme, can be used for it in recent years
Biosensor.But we not yet have found IrO at present2With Mimicry enzyme.
Sarcosine（Sarcosine）Also known as N methylglycine, be choline natural metabolism be glycine during one
Undoded amino acid intermediate can be reacted with methylamine by monoxone and is made, and can also pass through N- transmethylase by glycine
Effect generates.Sarcosine is pleasantly sweet, is dissolved in water, is present in the muscle and some other tissue of human body, normal under physiological status
Content of sarcosine in human serum is 1.59 ± 1.08 nmol/L.The choline or methionine that sarcosine can be taken in from diet
Metabolism, but be converted to glycine in vivo quickly.Glycine plays in person's physiological processes as structural amino acid
Important function, be the metabolic source of the living cells essential component such as glutathione, creatine, purine and serine.Sarcosine can swash
Prostate gland cancer cell living, and it is detected the generation for meaning malignant prostate cancer in urine.Sarcosine becomes forefront
The metabolite dramatically increased when gland cancer progress and transfer, can be detected in urine, thus be confirmed as forefront
One specific index of gland cancer.Therefore the detection of sarcosine plays a significant role in column gland cancer before the treatment.Common flesh ammonia
The detection method of acid has Electrochemiluminescprocess process, mass spectrography, fluorimetric analysis.Currently, having not been reported based on nano enzyme
Sarcosine detection method.Sarcosine oxidase（Sarcosine oxidase, abbreviation SOX, EC.1.5.3.1）Belong to flavoprotein
Oxidases can be catalyzed the redox of N methyl in sarcosine using FAD as co-factor, be creatinine in detection serum or urine
One of key enzyme of content is widely used in medical diagnostic field.It can be catalyzed sarcosine and be degraded to glycine, formaldehyde and peroxide
Change hydrogen.As a kind of important diagnosis enzyme preparation, SOX is widely used in the detection of creatinine level in human serum, for sentencing
The health degree of disconnected renal function.
In conclusion up to now, the sarcosine detection and analysis based on nano enzyme are rarely reported, IrO is had not been reported2Nanometer
The synthesis and its application of enzyme.
Summary of the invention
The purpose of the present invention is determine IrO2Nano enzyme synthesis and its as imitative enzyme material application.
To achieve the above object, the technical solution adopted by the present invention is：
A kind of nano enzyme material and its application, the IrO2Nano enzyme has imitative peroxidase activity.
Preferably, the imitative peroxidase material IrO2Nano enzyme synthesis process is：
(1) by K2IrCl6It is added in hydrogen citrate sodium solution, with NaOH tune pH to 7.5；
(2) flow back 30 min, after room temperature is cooling, then adjusts pH to 7.5, continues to flow back, and repeats the process until pH constant 7.5；
(3) solution is flowed back 2 hours further by bubble oxygen to get IrO2Nano enzyme.
It is further preferred, the imitative peroxidase material IrO2Nano enzyme can be used as imitative Catalyzed Synthesis By Peroxidase agent,
For the application based on imitative peroxidase activity.
More preferably, the imitative peroxidase material IrO2Nano enzyme is in acid condition as with peroxidase
Active catalyst, for carrying out qualitative/quantitative detection to sarcosine.Specific sarcosine detection method：
(1) phosphate buffer comprising certain density sarcosine, sarcosine oxidase（pH 8.0, 10 mmol/L）37
It is reacted 30 minutes at DEG C；
(2) above-mentioned solution is centrifuged, takes 10 μ L of supernatant, IrO described in claim 1-3 is added2Nano enzyme, organic chromogenic
Agent, acetate buffer（pH 3.5, 100 mmol/L）, continue to react 30 minutes at 37 DEG C；
(3) variation of observation solution colour is to realize qualitative detection；
(4) quantitative detection is realized using the corresponding light absorption value of microplate reader detection organicvisualization reagent oxide.
Organicvisualization reagent is added when the detection sarcosine；
The organicvisualization reagent is that 2,2 '-connection nitrogen-are bis-（3- ethyl benzo thiazole phenanthroline -6- sulfonic acid）Diamine salts（ABTS）Or 3,
3 ', 5,5 '-tetramethyl benzidines（TMB）.
Effect of the invention is：
1. the present invention has synthesized the IrO with peroxidase activity2Nano enzyme, and optimal pH is 3.5 or so, in acid item
Under part（pH 3.5−6.0）Show imitative peroxidase activity.
2. of the invention by IrO2Nano enzyme nanoparticle is applied to the qualitative quantitative analysis of sarcosine, realizes in acidity
Under the conditions of pH（pH 3.5−6.0）It is detected using qualitative/quantitative of the spectrophotometry to sarcosine.
3. the present invention provides the material for playing Mimicry enzyme under condition of acidic pH, it is applied to creatinine level in human serum
Detection be widely used in medical diagnostic field for judging the health degree of renal function.
Detailed description of the invention
Fig. 1 is IrO provided in an embodiment of the present invention2The transmission electron microscope figure of nano enzyme；
Fig. 2 is IrO provided in an embodiment of the present invention2Nano enzyme Mimicry enzyme effect picture；
Fig. 3 is IrO provided in an embodiment of the present invention2The pH effect of optimization figure of nano enzyme Mimicry enzyme；
Fig. 4 is H provided in an embodiment of the present invention2O2Quantitative measurement standard working curve；
Fig. 5 is the detection schematic diagram of sarcosine provided in an embodiment of the present invention；
Fig. 6 is the qualitative detection photo of sarcosine provided in an embodiment of the present invention；
Fig. 7 is the quantitative measurement standard working curve of sarcosine provided in an embodiment of the present invention.
Specific embodiment
Illustrate the contents of the present invention deeper into ground to become apparent from, will further enumerate some embodiments, but this hair below
It is bright to be not limited to cited embodiment.Specific experiment condition or method are such as not specified in the following example, by this field
Normal condition or method carry out.
Embodiment 1
Imitative peroxidase material IrO2The preparation of nano enzyme：
By 30 mg K2IrCl6It is added in 50 mL natrium hydrocitricums (3.8 mol/L) solution.Use NaOH（0.25 mol/
L）Acquired solution pH is adjusted to 7.5 by solution, is then flowed back 30 minutes under constant stirring.Hereafter, it is cooled down at room temperature, so
PH adjusting, stirring and reflux are carried out afterwards, repeat the process until obtaining constant pH 7.5.In order to obtain IrO2Nano enzyme hangs
Solution is further flowed back 2 hours by bubble oxygen, obtains IrO by supernatant liquid2Nano enzyme.Fig. 1 is to be seen by transmission electron microscope
The IrO observed2The TEM of nano enzyme is imaged.
Embodiment 2
IrO2The Mimicry enzyme of nano enzyme is verified：
Nano enzyme experimental system a：Catalystic converter system is to include H2O2（0.5 mmol/L）, above-described embodiment obtain IrO2It receives
Rice enzyme（70 µg/mL）, organicvisualization reagent TMB（0.5 mmol/L）Acetate buffer（pH 3.5, 100 mmol/L）.?
Room temperature（25 ℃）After lower reaction 30 minutes, the light absorption value in its 300 800 nm is detected using microplate reader.
Separately do two check experiments：IrO is not added in the catalystic converter system of one of control experiment b2Nano enzyme, with
Light absorption value is detected after reacting 30 minutes under above-mentioned experimental system similarity condition；The catalystic converter system of another control experiment c is
IrO2Nano enzyme（70 µg/mL）In acetate buffer（pH 3.5, 100 mmol/L）In, same as above-mentioned experimental system
Under the conditions of stand 30 minutes after detect light absorption value.
As shown in Fig. 2, experimental system a shows apparent peak, illustrate IrO2Nano enzyme has apparent at pH 4.0
The activity of imitative peroxidase；Check experiment b near 650 nm without apparent peak, if illustrating without IrO2Nano enzyme makees catalyst
It will be without significant reaction；Check experiment c, without apparent peak, illustrates that the peak of experimental system a is not IrO near 650 nm2Nano enzyme
The response peak of itself.
Embodiment 3
IrO2The pH of nano enzyme Mimicry enzyme optimizes：
Catalystic converter system is to include H2O2（0.5 mmol/L）,IrO2Nano enzyme（70 µg/mL）, organicvisualization reagent TMB（0.5
mmol/L）Different pH buffer（PH 1.0 2.0, glycine-HCI buffer；PH 3.0 6.0, Acetic acid-sodium acetate are slow
Fliud flushing；PH 6.5 8.0, phosphate buffer；PH 9.0 10.0, Tris- hydrochloride buffer；PH 11.0 12.0, bicarbonate
Sodium-sodium hydrate buffer solution）.In room temperature（25 ℃）After lower reaction 30 minutes, the extinction in its 650 nm is detected using microplate reader
Value.As shown in figure 3, IrO2Nano enzyme is at acid pH（pH 3.5−6.0）The activity of peroxidase is shown, and most
Suitable pH is 3.5 or so.
Embodiment 4
H2O2Quantitative detection：
Catalystic converter system is the H comprising various concentration2O2（0.01−3 mmol/L）,IrO2Nano enzyme（70 µg/mL）, it is organic
Color developing agent TMB（0.5 mmol/L）Acetate buffer（pH 3.5, 100 mmol/L）.In room temperature（25 ℃）Lower reaction 10
After minute, the light absorption value in its 300 800 nm is detected using microplate reader.It is drawn after deducting blank control by 650 nm light absorption values
Produce H2O2Standard working curve.As shown in figure 4,0 1.5 mmol/L of the range of linearity, linear equation is y=0.47x+0.15 (R2
=0.997)。
Embodiment 5
The qualitative detection of sarcosine：
Since sarcosine generates glycine, H under sarcosine oxidase catalysis2O2And HCHO, there is imitative peroxidase activity
IrO2Nano enzyme can be in H2O2In the presence of oxidation TMB colour developing, to realize the quantitative detection of sarcosine.Fig. 5 is the inspection of sarcosine
It measures and is intended to.Catalystic converter system is the sarcosine comprising various concentration（0−100 µg/mL）, sarcosine oxidase（300 µg/
mL）Phosphate buffer（8.0,10 mmol/L of pH）.After being reacted 30 minutes at 37 DEG C, above-mentioned solution is centrifuged, is taken
IrO is added in 10 μ L of clear liquid2Nano enzyme（70 µg/mL）, organicvisualization reagent TMB（0.5 mmol/L）, acetate buffer（pH
3.5,100 mmol/L）.10 min are reacted at 37 DEG C, as shown in fig. 6, observation color change.
Embodiment 6
The quantitative detection of sarcosine：
Catalystic converter system is the sarcosine comprising various concentration（0−100 µg/mL）, sarcosine oxidase（300 µg/mL）'s
Phosphate buffer（8.0,10 mmol/L of pH）.After being reacted 30 minutes at 37 DEG C, above-mentioned solution is centrifuged, supernatant is taken
IrO is added in 10 μ L2Nano enzyme（70 µg/mL）, organicvisualization reagent TMB（0.5 mmol/L）, acetate buffer（pH
3.5, 100 mmol/L）, after reaction 30 minutes, the light absorption value at its 650 nm is detected using microplate reader and draws out sarcosine
Standard working curve.As shown in fig. 7,0 60 μ g/mL of the range of linearity, linear equation is y=0.008x+0.012 (R2=
0.995)。
Claims (5)
1. a kind of iridium dioxide（IrO2）Nano enzyme, it is characterised in that：The IrO2Nano enzyme has imitative peroxidase activity.
2. iridium dioxide according to claim 1（IrO2）Nano enzyme, which is characterized in that synthesis step is as follows：
(1) by K2IrCl6It is added in hydrogen citrate sodium solution, with NaOH tune pH to 7.5；
(2) flow back 30 min, after room temperature is cooling, then adjusts pH to 7.5, continues to flow back, and repeats the process until pH is constant 7.5；
(3) solution is flowed back 2 hours further by bubble oxygen to get IrO2Nano enzyme.
3. one kind is based on IrO2The application of nano enzyme, it is characterised in that：IrO described in claim 1,22Nano enzyme can be used as
Imitative Catalyzed Synthesis By Peroxidase agent, for the application based on imitative peroxidase activity.
4. one kind is based on IrO2The sarcosine qualitative quantitative analysis application of nano enzyme, it is characterised in that qualitative quantitative analysis application
The step of it is as follows：
(1) phosphate buffer comprising certain density sarcosine, sarcosine oxidase（pH 8.0, 10 mmol/L）37
It is reacted 30 minutes at DEG C；
(2) above-mentioned solution is centrifuged, takes 10 μ L of supernatant, IrO described in claim 1-3 is added2Nano enzyme, organic chromogenic
Agent, acetate buffer（3.5,100 mmol/L of pH）, continue to react 30 minutes at 37 DEG C；
(3) variation of observation solution colour is to realize qualitative detection；
(4) quantitative detection is realized using the corresponding light absorption value of microplate reader detection organicvisualization reagent oxide.
5. organicvisualization reagent according to claim 4 is 3,3,5,5 '-tetramethyl benzidines（TMB）Or 2,2 '-connection
Nitrogen-is bis-（3- ethyl benzo thiazole phenanthroline -6- sulfonic acid）Diamine salts（ABTS）.
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| CN113466189A (en) * | 2021-05-25 | 2021-10-01 | 青岛农业大学 | Malathion colorimetric detection method based on double-enzyme activity inhibition effect |
| CN113466189B (en) * | 2021-05-25 | 2024-03-08 | 青岛农业大学 | Malathion colorimetric detection method based on double enzyme activity inhibition effect |
| CN113499773A (en) * | 2021-07-08 | 2021-10-15 | 辽宁大学 | Nano enzyme of nano zinc oxide supported palladium nanoparticles and preparation method and application thereof |
| CN114184789A (en) * | 2021-12-23 | 2022-03-15 | 云南大学 | Prostate specific antigen detection probe and prostate specific antigen detection kit |
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